臺灣博碩士論文加值系統

根據過去研究顯示：鉀離子通道與多種細胞包括腫瘤細胞株的增殖有關，以生長因子刺激T淋巴球以及上皮細胞都將使得此通道的表現量增加，且提升此通道活性亦會使增殖速率升高。另外，利用具有特異性的鉀離子通道阻斷劑，如tetraethylammonium (TEA)、4-aminopyridine(4-AP)、及quinine等藥物，會使得受生長因子刺激後的細胞增殖受抑制。此外，在腫瘤細胞中，鉀離子通道活性以及表現量的增加已被報告過。
根據衛生署九十年統計資料指出：口腔鱗狀上皮癌位居台灣男性十大癌症死亡率之第五位，且罹患人數及死亡率有逐年增加之趨勢。因此瞭解口腔癌化之基因異常，對於瞭解及控制口腔鱗狀上皮癌將有實質幫助。本實驗室過去用展示差異(differential display)之基因研究，發現鉀離子通道Kv 3.4在口腔癌細胞中有異常過度表現之現象。本研究中利用載體將Kv3.4基因片段轉染進入口腔鱗狀上皮癌細胞株OEC-M1，使其表現量增加，藉以探討此基因在口腔癌化過程中所扮演的角色。本研究結果中發現：Kv3.4的增加，有助於細胞生長的速度，但利用流式細胞儀來偵測，並無法發現細胞週期有明顯的變化。另一方面，Kv3.4的表現，有增強癌細胞抗藥性的現象，惟Kv3.4不會造成3T3細胞的轉型，可能不具有直接的致癌潛能。但其於癌症生成所扮演之角色仍值得深入探討。

Potassium channel have been reported to be involved in the proliferation of many types of cells, including tumor cell lines. Mitogenic stimulation increases the level of expression of potassium channel in T lymphocytes and epithelial cells. In addition, drugs that block potassium channels inhibit the proliferation of mitogen-stimulated normal human T lymphocytes, malignant rat lymphoma cells, human melanoma cells and human breast cancer cells. The over expression of potassium channel and related activity have been observed in neoplasms. Oral cancer is the fifth most common malignancy of the male population in Taiwan and more than 90 % of that are oral squamous cell carcinoma(OSCC) and the incidence and mortality of OSCC are increasing year by year. Our previous studies have shown that the abnormal expression of a voltage-gated potassium channel, Kv3.4 in an OSCC cell line by differential display. Preliminary studies also indicated the differential expression of Kv3.4 in normal appealing oral tissue and OSCC both in mRNA and protein level. We constructed the expression vector carring the Kv3.4 coding sequence, transfected into the OSCC cell line OEC-M1 and determined the high expression of Kv3.4 for subsequent carcinogenic studies. Our results suggest that the increasing expression of Kv3.4 may be beneficial for the cell growth without altering the cell cycle pattern of cancer cells. In addition, over expressing of Kv3.4 increased the chemoreistance of cancer cells. Although, Kv3.4 can not transform the 3T3 cells excluded the direct contribution of Kv3.4 in cell transformation, The potential role of Kv3.4 in carcinogenesis deserved further investigation.

目錄
目錄 ………………………………………………….. Ⅰ
表次目錄 …………………………………………….. Ⅱ
圖次目錄 ……………………………………………. Ⅲ
中文摘要 ……………………………………………. 1
英文摘要 ……………………………………………. 3
緒論 …………………………………………………. 5
材料與方法 …………………………………………. 15
結果 …………………………………………………. 29
討論 …………………………………………………. 33
表列 …………………………………………………. 37
圖列 …………………………………………………. 38
參考資料 ……………………………………………. 50
表次目錄
表一、核酸引子 …………………………………….. 37
圖次目錄
附圖一、電位控制與流入性整流器鉀通道結構圖………… 38
附圖二、電位控制性鉀通道之數狀分類圖………………… 39
附圖三、載體pGEM-T Easy及pcDNA輿圖……………… 40
附圖四、表現載體製備流程圖……………………………… 41
圖一、以RT-PCR偵測Kv3.4在OSCC及ESCC之表現… 42
圖二、構築表現載體之確認……………………………….... 43
圖三、偵測細胞之DNA所攜帶Kv3.4拷貝含量…………… 44
圖五、偵測細胞之Kv3.4之mRNA表現量……………...… 45
圖六、以MTT方式測得細胞之生長曲線…….……….…….. 46
圖七、以MTT方式測得細胞經藥物處理之存活率…….……. 47
圖八、3T3細胞送入Kv3.4後之比較………………….…… 48
圖九、細胞在電生理上的表現…………………………….… 49

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